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Clinical and Diagnostic Laboratory Immunology, July 1998, p. 531-536, Vol. 5, No. 4
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Development of a Human Gamma Interferon Enzyme Immunoassay and Comparison with Tuberculin Skin Testing for Detection of Mycobacterium tuberculosis Infection

Nuket Desem and Stephen L. Jones*

Biosciences Division, CSL Limited, Parkville 3052, Victoria, Australia

Received 12 November 1997/Returned for modification 5 February 1998/Accepted 29 April 1998

A sensitive two-step simultaneous enzyme immunoassay (EIA) for human gamma interferon (IFN-gamma ) has been developed and used as an in vitro test for human tuberculosis (TB) in comparison with tuberculin skin testing. The EIA was shown to be highly sensitive, detecting less than 0.5 IU of recombinant human IFN-gamma per ml within a linear detection range of 0.5 to 150 IU/ml. The assay was highly reproducible and specific for native IFN-gamma . In addition, the assay detected chimpanzee, orangutan, gibbon, and squirrel monkey IFN-gamma s. Cross-reactions with other human cytokines or with IFN-gamma s derived from mice, cattle, or Old World monkeys were not evident. The assay was used to detect TB infection by incubating whole blood overnight with human, avian, and bovine tuberculin purified protein derivatives (PPDs), as well as positive (mitogen)- and negative-control preparations. The levels of IFN-gamma in plasma supernatants were then determined. Blood from 10 tuberculin skin test-positive individuals responded predominantly to the human tuberculin PPD antigen and to a lesser extent to bovine and avian PPD antigens. By contrast, blood from 10 skin test-negative individuals showed minimal responses or no response to any of the tuberculin PPDs. Detectable levels of IFN-gamma were present in all blood samples stimulated with mitogen. In vivo tuberculin reactivity was correlated with IFN-gamma responsiveness in vitro. These results support the further study of the blood culture-IFN-gamma EIA system as an alternative to skin testing for the detection of human TB infection.

* Corresponding author. Mailing address: Biosciences Division, CSL Limited, 45 Poplar Rd., Parkville 3052, Victoria, Australia. Phone: 61 3 9389 1778. Fax: 61 3 9389 1224. E-mail: sjones{at}csl.com.au.

Clinical and Diagnostic Laboratory Immunology, July 1998, p. 531-536, Vol. 5, No. 4
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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